Tags

Type your tag names separated by a space and hit enter

Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures on Collagen and EHS Sarcoma Matrices.
PLoS One. 2015; 10(4):e0124867.Plos

Abstract

Liver-enriched transcription factors (LETF) play a crucial role in the control of liver-specific gene expression and for hepatocytes to retain their molecular and cellular functions complex interactions with extra cellular matrix (ECM) are required However, during cell isolation ECM interactions are disrupted and for hepatocytes to regain metabolic competency cells are cultured on ECM substrata. The regulation of LETFs in hepatocytes cultured on different ECM has not been studied in detail. We therefore compared two common sources of ECM and evaluated cellular morphology and hepatocyte differentiation by investigating DNA binding activity of LETFs at gene specific promoters and marker genes of hepatic metabolism. Furthermore, we studied testosterone metabolism and albumin synthesis to assess the metabolic competence of cell cultures. Despite significant difference in morphological appearance and except for HNF1β (p<0.001) most LETFs and several of their target genes did not differ in transcript expression after Bonferroni adjustment when cultured on collagen or Matrigel. Nonetheless, Western blotting revealed HNF1β, HNF3α, HNF3γ, HNF4α, HNF6 and the smaller subunits of C/EBPα and C/EBPβ to be more abundant on Matrigel cultured cells. Likewise, DNA binding activity of HNF3α, HNF3β, HNF4α, HNF6 and gene expression of hepatic lineage markers were increased on Matrigel cultured hepatocytes. To further investigate hepatic gene regulation, the effects of Aroclor 1254 treatment, e.g. a potent inducer of xenobiotic defense were studied in vivo and in vitro. The gene expression of C/EBP-α increased in rat liver and hepatocytes cultured on collagen and this treatment induced DNA binding activity of HNF4α, C/EBPα and C/EBPβ and gene expression of CYP1A1 and CYP1A2 in vivo and in vitro. Taken collectively, two sources of ECM greatly affected hepatocyte morphology, activity of liver enriched transcription factors, hepatic gene expression and metabolic competency that should be considered when used in cell biology studies and drug toxicity testing.

Authors+Show Affiliations

Centre for Pharmacology and Toxicology, Hannover Medical School, Hannover, Germany.Centre for Pharmacology and Toxicology, Hannover Medical School, Hannover, Germany.Centre for Pharmacology and Toxicology, Hannover Medical School, Hannover, Germany.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25901575

Citation

Borlak, Jürgen, et al. "Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures On Collagen and EHS Sarcoma Matrices." PloS One, vol. 10, no. 4, 2015, pp. e0124867.
Borlak J, Singh PK, Rittelmeyer I. Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures on Collagen and EHS Sarcoma Matrices. PLoS One. 2015;10(4):e0124867.
Borlak, J., Singh, P. K., & Rittelmeyer, I. (2015). Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures on Collagen and EHS Sarcoma Matrices. PloS One, 10(4), e0124867. https://doi.org/10.1371/journal.pone.0124867
Borlak J, Singh PK, Rittelmeyer I. Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures On Collagen and EHS Sarcoma Matrices. PLoS One. 2015;10(4):e0124867. PubMed PMID: 25901575.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures on Collagen and EHS Sarcoma Matrices. AU - Borlak,Jürgen, AU - Singh,Prafull Kumar, AU - Rittelmeyer,Ina, Y1 - 2015/04/22/ PY - 2014/08/07/received PY - 2015/03/13/accepted PY - 2015/4/23/entrez PY - 2015/4/23/pubmed PY - 2016/4/5/medline SP - e0124867 EP - e0124867 JF - PloS one JO - PLoS One VL - 10 IS - 4 N2 - Liver-enriched transcription factors (LETF) play a crucial role in the control of liver-specific gene expression and for hepatocytes to retain their molecular and cellular functions complex interactions with extra cellular matrix (ECM) are required However, during cell isolation ECM interactions are disrupted and for hepatocytes to regain metabolic competency cells are cultured on ECM substrata. The regulation of LETFs in hepatocytes cultured on different ECM has not been studied in detail. We therefore compared two common sources of ECM and evaluated cellular morphology and hepatocyte differentiation by investigating DNA binding activity of LETFs at gene specific promoters and marker genes of hepatic metabolism. Furthermore, we studied testosterone metabolism and albumin synthesis to assess the metabolic competence of cell cultures. Despite significant difference in morphological appearance and except for HNF1β (p<0.001) most LETFs and several of their target genes did not differ in transcript expression after Bonferroni adjustment when cultured on collagen or Matrigel. Nonetheless, Western blotting revealed HNF1β, HNF3α, HNF3γ, HNF4α, HNF6 and the smaller subunits of C/EBPα and C/EBPβ to be more abundant on Matrigel cultured cells. Likewise, DNA binding activity of HNF3α, HNF3β, HNF4α, HNF6 and gene expression of hepatic lineage markers were increased on Matrigel cultured hepatocytes. To further investigate hepatic gene regulation, the effects of Aroclor 1254 treatment, e.g. a potent inducer of xenobiotic defense were studied in vivo and in vitro. The gene expression of C/EBP-α increased in rat liver and hepatocytes cultured on collagen and this treatment induced DNA binding activity of HNF4α, C/EBPα and C/EBPβ and gene expression of CYP1A1 and CYP1A2 in vivo and in vitro. Taken collectively, two sources of ECM greatly affected hepatocyte morphology, activity of liver enriched transcription factors, hepatic gene expression and metabolic competency that should be considered when used in cell biology studies and drug toxicity testing. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/25901575/Regulation_of_Liver_Enriched_Transcription_Factors_in_Rat_Hepatocytes_Cultures_on_Collagen_and_EHS_Sarcoma_Matrices_ L2 - https://dx.plos.org/10.1371/journal.pone.0124867 DB - PRIME DP - Unbound Medicine ER -